This application builds on the advances that the USC investigators have made in nanoscale materials and devices, including nanoscale field effect transistors (FETs), microfluidics and biosensors. It has become increasingly clear that it is critical to develop technology platforms that permit quantitative analysis of multiple serum protein biomarkers as well as multiple genetic and epigenetic lesions in a cost-effective manner for early detection, prognostic evaluation and therapeutic monitoring of cancer. To address this critical public health issue, the overall goal of this project is to develop and validate novel nanowire (NW) and single-walled carbon nanotube (SWNT) sensor arrays for single-step detection of serum protein markers, and of DNA mutation, polymorphism and site-specific methylation. This application is focused on the development of multiplexed nanosensor arrays that can be used in a clinical setting. We have demonstrated that In2O3 NW and SWNT- based FETs can be used as effective biosensors with a potential to achieve both high specificity and sensitivity. We propose to develop and test the applicability of the NW/NT FET arrays for multiplexed sensing in model systems as well as in clinical serum samples. Unique surface functionalization for various capture ligands will be achieved via our proven microfluidic delivery system, and semiconductor processing will be employed to create the proposed biosensors. In Specific Aims 1 to 3, we will used novel NW/SWNT sensors to detect 1) cancer related protein biomarkers (including PSA, CEA, CA-125 and other markers as the model);2) DNA nucleic acid sequence changes related to cancer (using K-ras mutations as the model);and 3) site- specific DNA methylation (epigenetic) cancer related changes (using p16 gene as the model). In Specific Aims 4 and 5, we will integrate a novel microfluidic delivery and test system and develop nanosensors arrays capable of interrogating multiple biomarkers simultaneously. Both NW and SWNT-based devices will be integrated side-by-side into the same array, a powerful design which will allow complementarily of signals for analyte binding, minimizing false negative or positive signals for complex mixtures. The sensitivity, selectivity, stability and reproducibility of these sensor arrays will be investigated. In Specific Aim 6, we will test serum from cancer patients with known levels of relevant biomarkers. The applications addressed here will result in sensor array platforms that are highly likely to yield label-free, low-cost, multiplexed assays for molecular diagnosis and monitoring therapeutic efficacy for cancer, and which can easily be extended to applications to other diseases as well. Project Narrative/Relevance: It is becoming increasingly clear that in order to achieve the goal of early detection and therapeutic monitoring of cancer using serum based biomarkers, including proteins, DNA mutations, and other changes in DNA, it is critical to develop technology platforms that permit quantitative analysis of multiple biomarkers simultaneously and a cost effective manner. In order to address this critical public health issue, the overall goal of this project is to develop and validate novel nanowire (NW) and single-walled carbon nanotube (SWNT) sensor arrays for single step detection of serum based protein and DNA biomarkers. The research in this application is highly likely to result in novel devices that will allow for the detection of multiple cancer associated biomarkers capable of use in early detection and therapeutic monitoring of cancer. It is also likely that this device will be easily extended to applications in other diseases as well.